IntroductionPVDF membrane was originally introduced to protein use as an ideal
medium for the harsh chemical environment of N-terminal, or Edman degradation,
sequencing. Even though PVDF is very hydrophobic and requires a prewetting
step in alcohol, its high protein binding capacity, target retention,
and resistance to cracking have made it an appealing membrane for general
laboratory techniques.

The two main applications for PVDF are N-terminal sequencing and immunoblotting,
both of which benefit from the qualities of PVDF but rely on different
features of the membrane. While sequencing work is concerned with retaining
as much protein as possible, a western blot requires good signal retention
with very low background. To provide the best membrane for each technique,
Bio-Rad offers two grades of PVDF: Immun-Blot PVDF membrane for western
blotting and Sequi-Blot PVDF membrane for protein sequencing.

Immun-Blot PVDF Membrane
The focus with a blotting membrane is on how well it delivers signal while
resisting background and nonspecific binding. Immun-Blot PVDF membrane
is ideal for chemiluminescent (Figure 1) and colorimetric (Figure 2) western
blots because it very strongly retains target protein (140150 g protein/cm2
membrane) but resists background that can obscure highsensitivity detection.
Immun-Blot PVDF membrane retains proteins in any transfer format: tank
blotting, semi-dry blotting, and dot blotting all deliver excellent results.
For proteins that are difficult to transfer, up to 0.1% SDS can be added
to the transfer buffer without affecting the binding of the proteins to
PVDF. The results are consistently clean, easy-to-read blots. The physical
strength of Immun-Blot PVDF membrane means that it will not crack or tear
during common handling, and will hold up under repeated stripping and
reprobing applications.

Sequi-Blot PVDF Membrane
For protein sequencing applications, Sequi-Blot PVDF membrane is the best
choice due to its extremely high protein binding capacity of 170200 g/cm2.
This is the original Bio-Rad PVDF membrane, designed to withstand the
conditions of N-terminal sequencing while providing the binding capacity
to sequence even low-abundance samples. The table shows the increased
protein recovery possible with Sequi-Blot PVDF membrane. Recovery of blotted
proteins is typically in the range of 80100% of the initial sample load,
resulting in higher initial coupling yields.

The binding efficiency of Sequi-Blot PVDF membrane is illustrated in
Figure 3. This experiment showed that Sequi-Blot PVDF membrane is able
to retain proteins that transfer through a competitors product.

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